Ethnobotanical inventory and medicinal perspectives of herbal flora of Shiwalik mountainous range of District Bhimber, Azad Jammu and Kashmir, Pakistan

The present study was carried out to document traditional ethnobotanical knowledge (TEK) of herbal flora of District Bhimber, Azad Jammu and Kashmir (AJK), Pakistan to explore medicinal potential of wild indigenous plants (WIPs) for drug discovery. The research data was conducted during years 2015–2017 using questionnaire proforma employing structured and semi-structured interview models. The informants belonged from indigenous communities comprising of both genders with age ranging from 35–105 years. The interviews were conducted using local translator or guide who knew the dialects of all local languages. The study produced ethnobotanical inventory of 173 herbal species belonging to 45 families and out of these Poaceae was dominant family with 27 species. It was explored that maximum herbal species depicted multi-usage especially food, fodder and fuel. Among 173 herbal plant species, 69% species were used as fodder, 72% species as fuel source, 9.8% as ethnoveterinary medicines, 16% for home construction, 12% for cosmetics, 5.2% as honeybee plants and 2.7% were used as fiber source by the local people. Many local plants have been in promulgation for cure of different diseases response of the interviewees recorded during survey in correlation with authenticating of traditional data. The RFC values represented the relative popularity of individual species in study area according to their use values. The highest value was calculated for Alternanthera pungens L. (0.90) and followed by Achyranthes aspera L. (0.80. The study reveals that many species are known for commonly used in traditional ethnomedicines (TEMs). Due to different biotic and abiotic factors in conjunction with climatic changes many herbal flora of Shiwalik mountain range (SMR) of District Bhimber of AJK is under threat. The factors like habitat loss, overgrazing, construction of communication infrastructure, silviculture prac-tices, shelter construction (houses) and other more domestic use of wild land by clearing wild lands are boosting towards plant biodiversity loss. There is dare need to work on com-prehensive exploration of TEMs to discover neo drugs from wild indigenous plants and do work for conservation of wild flora of the area for future generations.


Introduction
Plant resources provide all necessary life sustenance materials like food, fodder, feed, forage, shelter, medicines and aesthetic values for people an area [1]. It is worth to state that a little work has been conducted on determination of conservation status of wild flora of many areas of the world. Old people or herbalists (hakims) are connected with medicinal plants and use different herbs for preparing medicines which are promulgated in the society [2,3]. Indigenous people of rural areas of the world are primarily dependent on wild resources for fulfilling their daily life requirements. Plants play a central role in the chief health care services to the inhabitant of the area. They provide healing agents and significant raw materials to built-up of old and new medicines of homeopathic and allopathic form [3,4]. About 80% world population depends on plants for health issues and to treat different infirmities. Herbal flora plays pivotal role in provision of nutritive and food subsistence to the dwellers of the mountains and rural areas of the world. The herbaceous plants are also key source of fodder and forage for the domesticated animals and rodents as well as for wild fauna.
Azad Jammu and Kashmir (AJK) area is rich in floral diversity because of the diverse habitats, such as streams, rivers, meadows, lakes, springs, waste lands, slopes, cultivated fields, etc. The present study was designed to study the floristic and ethnobotanical uses of herbal flora by indigenous communities of Shiwalik mountaineous range (SMR) of District Bhimber Azad Jammu and Kashmir, Pakistan (Fig 1). The study area falls in District Bhimber which was declared as an independent District in year 1996 with an area is 1516 Km 2 .
This area is also called as 'Gateway to Kashmir (Bab-e-Kashmir), as Mughal emperors and warriors entered through this region to Kashmir valley and subcontinent [3,5]. Geographically Bhimber valley is situated on 32˚58'11"N Latitude and 74˚4'11"E longitudes with its edaphalogical conjunction with Gujrat, Jhelum areas of Punjab province and west side with KPK province of Pakistan while on north and east side different areas of Indian occupied Kashmir [3,6]. The rationale of the study was based on fact that "study comprises of mountains chain known as Shiwalik mountain range (SMR) which has rich phytodiversity and indigenous people of the area primarily depend wild plants and fauna resources for life sustenance. This area is yet to explore and this first time study conducted with a hypothesis that "the ethnobotanical study will assist in documentation and preservation of traditional culture of different ethnic tribes and exploration of novel botanic drug uses from medicinal plants of the area". The pivotal key objectives of the study were multifarious comprising of (i) document floristic and ethnobotanical inventory of wild herbal flora of SMR area, (ii) to prepare data of important traditional ethnomedicines (TEMs) and enlist their botanic recipes and (iii) to determine conservation status of different plants and recommend different propagation methods.

Study area
The study area comprises of hilly terrains of Shiwalik mountain range (SMR) of District Bhimber of AJK with a diverse topography (plains and hills) and phytogeography (diverse phytodiversity). The District Bhimber has mostly acidic and sandy-clay soil in nature with arid climate receiving scarce rainfall spells. Higher rainfall is recorded in July and August and lower in winter season. The average maximum and minimum temperature are 28.9˚C and 15.8˚C, respectively. The total average rainfall is about 1233 mm per year [7]. Average rainfall is 102.8mm per month. The area has temperature range between 0˚C to 40˚C with summer is bit hotter while at high altitude and forest zones the ranges preferably remains cool and low [8].

Protocols used for data collection
In this part of research, data was collected using semi-structured and structured interview protocols following standard procedure which will consist of questionnaire and field plant interview technique [3,9]. Many planned field trips were arranged and the area was visited with assistance of women guide or translator who was very useful to collect data from female communities of villages of SMR area of Bhimber. The interviewees were between age of 35 to 105 years comprises of both genders with diverse professions people but most were peasants and wood cutters. During Interviews using a questionnaire method to document the ethnobotanical knowledge on the herbal plant quantities, resources and their utilization by the farmers, elder people usually with the age of more than 70 years, drug dealers, hakims, shopkeepers etc. Plants were grouped on the basis of their fodder, honey bee plants, ethnoveterinary medicines, fuel and ornamental etc. The plant specimens collected, pressed, dried, preserved and were arranged alphabetically by family name, flowering period and vernacular name. The collected specimens were identified by comparing with floristic literature [10][11][12][13]) and online data comparison (www.theplantlist.org and www.eflora.org), following procedure of previous researcher [14].

Ethical statement
The permission for field study and collection of wild plants (herbs) from wild forest areas of Shiwalik Mountian Range (SMR) zones of District Bhimber (AJK) was obtained from Departmental ethical committee (DEC) on official letter (Ref No: 31/DEC/BOT/2015; Date: 20/06/2015) and counter signed by /Head of Department. For data collection from indigenous informants, a prior verbal permission was obtained from each family head/elder man and village leader. It as acknowledged that perosnal information of each interviewee will be kept confidential as per advice and rule of DEC.

Field survey permission letter
An official field permit (Ref No: DFO/655/2015 Dated: 01/07/2015) was obtained from District Forest Officer (DFO) to make field visits of forest area of SMR, DistrictBhimber of AJK and to collect herbal samples of plants. The right of obedience of forest rules was fully followed as per SOPs provided by the relevant office and guidance of DEC was also fully obeyed.

Quantitative ethnobotanical tools
Ethnobotanical (EB) data collected through questionnaire method was tabulated and analyzed using quantitative analytical tools to confirm its reliability and authenticity. Several statistical tools were used for concentrating data regarding EB from study area following procedure of Amjad et al., 2017 andIshtiaq et al., 2021 [3, 15]. For analysis of indigenous knowledge gathered from study area and its authentication was confirmed by using micro statistical tools like: "informant consensus factor" (ICF), "relative importance of plant" (RIP), "fidelity level" (FL), and "rank order of priority" (ROP) following the procedures of previous researchers like Ishtiaq et al., (2021) [3] and these quantitative ethnobotanical indices were prevalently utilized by previous researchers [16][17][18].
Informant Consensus Factor (ICF). The data collected from the respondents regarding the use of various plant species were legitimated and verified by using a tool named informant consensus factor (ICF). ICF was calculated from gathered questionnaire data by using following equation; where, Nur represents number of use citations minus (-) Nt total number of plant species used cited by respondents; divided (/) by the number of use citations in each category minus one as per given formula above. If the result is nearer to (1) or exactly (1) then reported plant species is abundantly used by locals in the area. Whereas, if value is near to (0) or comes out to be zero then reported plant species is casually used by inhabitants of area [19]. Fidelity Level (FL). The fidelity level of the data was calculated which depicts the percentage (%age) of interviewees claiming any use of a particular plant for the same major purpose or field and total number of commonly reported usages or ailments. It was calculated according to formula: Whereas; Np depicts number of informants, who claimed a particular use of a plant species used for a typical disease and N means that number of informants/interviewees who used the plants as an ethnomedicine to treat given diseases [3,20]. Rank Order Popularity (ROP). To determine correction between FL and RPL rank order popularity is used. ROP designate the popularity rank to individual in accordance to FL and RPL values [3,21]. Following formula was used for ROP calculation: ROP value represents higest rank/level for popularity of various MPs used to cure specific disorder RPL and ROP statistical test are significantly important for determining EB importance of commonly used plant in respected area.
Relative Popularity Level (RPL). RPL describes the frequency use of a particular species. In some analysis, sporadically FL provides same usage frequency of various species then Relative popularity level index is used to reassure and confirm FL values of various trial species. RPL value generally ranges in-between 0-1, where value 1 or nearer value to 1 represents maximum RPL value while 0 or nearer to 0 represents minimum value frequency for particular disease [3,22]. Value of RPL increases in accordance with informants as it rises correlation coefficient factor r = 0.10. For example, if a particular species is reported by 25 or more individuals it will have high RPL value and vice versa. RPL is calculated by proportion of (I u /number of individuals reported the use of particular plant). When 25 of more individuals report for a particular species RPL value becomes (I u /12) which gives one (1) and species is ranked as Popular (P). Whereas if less than 25 individuals report plants uses then respected plant species is ranked as unpopular (UP) as stated by Ishtiaq et al., [23,24]. Marginal value in-between, P and UP of particular plant refers point where further increase in number of informants don't upswing medicinal use per plant species.
Relative Importance of Plant (RIP). To determine the pharmacological or pharmaceutical significance of individual plant relative importance of plants (RIP) was used following protocol of Umair and Amjad [25]. where BS is the number of body System healed by single plant species whereas, rel. BS is total relative number of body system treated by the given plant species.

RIP
Use value index. The use value index (UVI) demarcates relative importance of different uses of the specific species. It was determined by using past cited formula following protocol of Leto et al., 2013 [26, 27].
Whereas UV indicates "relative use value" of the single species; "U" is the 'number of uses mentioned by each informant for the species and "N" is the 'total number of informants who reported that species.
Relative Frequency of Citation (RFC). Relative frequency of citation is used an index to explore significance or importance of each species occurring in the local area. RFC was determined by "dividing the number of informants" confirming the frequency.
Where, '0 < RFC >1'; and FC is the 'number of informants' reporting use of a particular species and N is the 'total number of informants' involved in study survey.
Priority Ranking (PR). Priority ranking (PR) method was used for indicating the preference of the local people about the potential use of each plant in certain use and it describes the biotic pressure on the plants of the area and it assists to calculate the conservation status of various species in the study area [28,29].

Results and discussion
The research work was carried out in District Bhimber during years 2015-2107 prevalently based on SMR areas of AJK, Pakistan. The plants samples free of disease were collected, pressed, dried in newspapers and mounted as per standard herbarium protocol of Ishtiaq et al., (2021) [3]. These specimens were mounted on herbarium sheets, then identified, preserved and deposited in Herbarium of Mirpur University of Science and Technology Bhimber Campus, AJK. In the investigated area 173 herbaceous plant species belonging to 45 families were found and recorded for EB and TEMs data. Among 45 families Poaceae were the dominant families with 27 species. There were followed by Papilionaceae, Amaranthaceae and Fabaceae with 8 and 7, 7 species, respectively. These were followed by Boraginaceae and Lamiaceae with 06 species, Cyperaceae 05 species each, Chenopodiaceae, Euphorbiaceae, Plantaginaceae, Ranunculaceae and Solanaceae 04 species each and Acanthaceae, Liliaceae, Malvaceae, Brassicaceae, Cucurbitaceae, Portulaceae, Papaveraceae 03 species each (Table 1). Similar results were explored and published by Ajaib et al., 2014 in District Kotli, Azad Jammu and Kashmir [30]. Plant is an unusual organism of infinite kindness and compassion it offers to mankind, the creation of its life activities in different ways. Among them domestic, commercial and industrial uses are well known.

Ethnobotanical findings
Ethnobotany includes all aspects of traditional uses of plants including food, poison, fodder, timber, clothing, dyes, fuel, medicines and veterinary medicines. Ethnobotanical survey of SMR region of District Bhimber gave evaluation of relation between man and his surrounding plants. For this purpose, many field surveys were carried out in the certain selected sites of the study area of SMR. Many ethnobotanical plant species were recognized along with data about traditional uses and villagers of the area. Among 173 herbal plant species, 69% species were used as fodder, 72% species were used for fuel purpose, 9.8% were used as ethnoveterinary purposes, 2.7% were used as fiber, 5.2% were used as honeybee plants, 16% were used as domestic and 12% were used as cosmetics (Table 2 and Fig 2). Similar studies were also conducted by Hussain et al., 2012 when they studied the ethnobotany of plants (Angiosperm) flora of Rawalakot. They represented 173 medicinal plants used by native people for different purposes and cure of human and livestock. About 111 species are weeds, 158 used for fodder, 68 species used as fuel, 33 species were fruit yielding 22 were used as timber, 31 as ornamental, 25 as fencing lawns and fields. While 21 species were used as vegetables, 9 for thatching houses and huts; and 13 species were recorded as poisonous. Shaukat et al., (2012), studied ethnobotanical studies of diversity of plants of some selected regions of Rawalakot [31]. They concluded that 26 plant species belonging to 19 families were used by local people for different purposes. Sedges and grasses of the study area were grazed as fodder by animals in Summer and Spring seasons and stored for Winter and Autumn when no fodder is accessible for grazing. The relationship between plants and people is extraordinary because plants provide fodder, domestic animals, food, ethno veterinary, fuel and honeybee, flowers for esthetics and happiness [30,32]. It was studied that due to grazing off domestic animals in the hilly areas of District Bhimber and over harvesting of every available portion of the gainful medicinal plants by indigenous communities as a source of income creates severe plethora of biotic pressure leading towards loss or extinction in future. Generally, women were mostly linked with collection of herbal plants as source of botanic drugs who collect and dry these phyto-drugs at home, on rocks or on mats, then after sprinkling the plants on roof. The dried collected herbal plants were sold directly to the main city markets and provincial drug traders. Similar results were also shown by Khan et al., (2011) during whose work conducted on the ethnomedicinal evaluation of plants of Hindu-Kush area of Pakistan [33].

Ethnomedicinal uses
Among173 herbaceous plants, belonging to 12 families, 21.4% species used against skin diseases followed by 16.7% used for wound healing and 14.4% used against fever. These were followed by 17.3% for stomach problems, 12.13% for cough and 22.7% for treatment of cancer.
The study revealed that17.14% species were used against jaundice, 4.6% were used against asthma and 8.09% for kidney diseases. These were followed 9.2% of herbal flora which was used for diabetes, 4.62% against snake biting and 1.7% species were used for treatment of tooth problems (Tables 3 and 4; Fig 2). Similar results were also shown by Shinwari, ( Fig 3). The similar results have been reported in earlier works that had been directed in Island and Italy which described that the leaves were frequently used in herbal medicines [39,40]. It has been found that extreme assembly of leaves along with other plant parts like bark and roots because leaves have more efficiency than other parts because leaf has a greater number of chemical ingredients [41].

Informant Consensus Factor (ICF) and Fidelity Level (FL)
The major aliments were classified into 12 disease categories by using ICF. According to ICF values the highest numbers of plant species were used against wound healing, snake bite, skin diseases, eye diseases and asthma (Table 6; Fig 4). Brassica campestris L. with maximum FL (40.0) followed by Aloe vera L. with FL (36.6%) and Adiantum capillus-veneris L. with FL (30.0%) were used as antidiabetic and skin diseases, arthritis, respectively (Table 7). Similar

MUH-1721
Field marigold Asteraceae Leaf and flowers Extract of leaves is used to cure antiseptic, cure skin diseases and healing of wounds.

MUH-1722
Distaff thistle Asteraceae Flowers, seeds The paste of seeds is prepared and applied to cure different skin diseases and fever. The flower decoction is used to fever and chest cold feelings.

MUH-1723
Marigold Asteraceae Leaf, flowers The sap of flowers used to cure skin diseases and treatment of cancer. The external use of plant in form of extract with tincture is effective for wound healing, blood purifier, cure skin disorders, anticancer, anemia and kidney diseases

35.
Carthamus oxyacantha L. Kandyari Asteraceae Flowers, seeds Flowers are used to cure jaundice while oil extracted from seeds is used to cure skin diseases and cure jaundice. MUH-1724
Mexicana tea Chenopodiaceae Leaves, flowers, and seeds.
Dried leaves are used to treat cancer. Seed oil is used to cure arthritis and skin diseases. MUH-1725

MUH-1726
Oak Chenopodiaceae Leaves and seeds Fresh leaves extract is used to cure kidney diseases, arthritis, skin diseases, digestive, cough and pectoral pains.

MUH-1727
Bathu Chenopodiaceae Leaves Leaf juice is used to cure kidney problems and cure spleen enlargement problems.       [3]. ICF results for ethnomedicines were fluke with past investigators available by different experts in respite of the world [42]. The ICF values of the diseases occurring in the area were similar with past studies conducted in different areas of Pakistan [43] and Azad Kashmir [36,44].

Relative Frequency Citation (RFC) and Use Value (UV) of ethnobotanical uses of herbs
Ethnobotanical significance of many herbs was calculated by using relative frequency of citation (RFC) and use value ( (Table 8). Similar findings were reported by the previous researchers who depicted that quantitative tools are very valuable in analysis of significance of each species. Coinciding results were provided in the past works who stated that these cited species of high medicinal value and can be used for drug discovery through analytical studies [45,46].    (Table 9). Similar findings have been reported in previous research work, where herbal has been predominantly used as source of medicines and other life sustenance materials [47][48][49].

Priority Ranking (PR)
According to the information gathered it is predicted that use of plant for various house cores is common in study area. Extensive use of plant and its products causes reduction in respected abundance of plant species. The priority ranking (PR) analysis determines the abundance status of each ethnobotanically reported plants species. Data was organized into tubular form and destructive order was determined and arranged in six Destructive order is; 4<3<2<1 = 4 number represents the most destructive value, as shown in Table 10. The results depicted that silviculture and clearing of land for agriculture purpose was the key issue of pressure on the herbal flora because due to this phenomenon herbs were cut off or made died due to chemical spray. The fire due to random or deliberate use by farmers to clear unwanted flora from the wild areas was the second important factor affecting the herbaceous flora of the study area.  Similar results have been cited by the previous researchers that herbal flora is very sensitive and fragile being easily diminished due to natural or anthropogenic activity [50][51][52].

Conservation status determination
The Inhabitants of the area relay on natural sources firstly due to lack of resources and secondly due to high cost of mark product. Alongside native inhabitant also believes that herbal medicines are safer for use as compare to pharmaceutical medicines. Extensive use of natural sources is causes serious risk to their abundance percentage. Calculated values represent classification of plants according to risk factors decreasing their abundances. Values determine how respondents of area categorize different threatening parameters for flora of study area. Highest rank of priority ranking (PR) is recorded for consumption of trees for fuel, followed by wood cutting of plants, while lowest is recorded for hedging and thatching. This shows that villagers of study area highly depend upon wood for furniture making, for fuel, in construction and tools making. Whereas fresh parts of trees are used for fodder. This represents key factor that will cause natural flora degradation in study area. Many species like Aerva sanguinolenta, Ajuga bracteosa, Boerhavia diffusa, Citrullus colocynthis, Hyoscyamus niger, Solanum surratense, Trichodesma indicum and Viola canescens are declared as threatened species and some of these are endangered. It species is near to extinct from the study area which demands urgent need to conserve these valuable plants. Similar efforts and findings have been cited and recommended for conservation of wild flora of in different areas of Pakistan and world [3,9,[53][54][55][56][57]. These plants are of high medicinal potential and conservation of these will provide commercial and drug development opportunity for the future generation.

Recommendations and future threats managements
There is dare need to tree flora of the area because it provides lot of ethnobotanical and folklore herbal therapeutics to cure different ailments in SRM area of District Bhimber, AJK. At a    strict rules at governmental level to conserve of most common EB plant species from degradation. There is need to expand the Prime ministers 'billion's tree tsunami' project by involvement of youth in its propagation, cultivation and protection. This will lead towards 'green revolution' and make the environment eco-friendly with least pollution impacts as well as timely raining for good agriculture products and crops.